Abstract
3-Chloro-1, 2-propanediol (3-MCPD) is a food-borne toxic substance well-known for more than 40 years that is mainly associated with nephrotoxicity. A better understanding of 3-MCPD nephrotoxicity is required to devise efficacious strategies to counteract its toxicity. In the present work, the role of endoplasmic reticulum (ER) stress along with its underlying regulatory mechanism in 3-MCPD-mediated renal cytotoxicity was investigated in vivo and in vitro. Our data indicated that 3-MCPD-stimulated ER stress response evidenced by sustained activation of PERK-ATF4-p-CHOP and IRE1 branches in Sprague Dawley (SD) rats and human embryonic kidney (HEK293) cells. Moreover, ER stress-associated specific apoptotic initiator, caspase 12, was over-expressed. Blocking ER stress with its antagonist, 4-phenylbutyric acid (4-PBA), improved the morphology and function of kidney effectively. 4-PBA also increased cell viability, relieved mitochondrial vacuolation, and inhibited cell apoptosis through regulating caspase-dependent intrinsic apoptosis pathways. Furthermore, the enhanced expressions of two mitochondrial fission proteins, DRP1/p-DRP1 and FIS1, and the relocation of DRP1 on mitochondria subjected to 3-MPCD were reversed by 4-PBA, while the expression of the fusion protein, MFN2, was restored. Moreover, cellular Ca2+ overload, the over-expression of CaMKK2, and the loss of mitochondria-associated membranes (MAM) were also relieved after 4-PBA co-treatment. Collectively, our data emphasized that ER stress plays critical role in 3-MCPD-mediated mitochondrial dysfunction and subsequent apoptosis as well as blockage of ER stress ameliorated kidney injury through improving mitochondrial fission/fusion and Ca2+ homeostasis. These findings provide a novel insight into the regulatory role of ER stress in 3-MCPD-associated nephropathy and a potential therapeutic strategy.
Graphical abstract
Graphical Headlights
1. 4-PBA inhibits ER stress mainly through regulating PERK-ATF4-CHOP and IRE1-XBP1s branches.
2. Inhibition of ER stress by 4-PBA mitigates ER associated and mitochondrial apoptosis
3. Inhibition of ER stress by 4-PBA helps maintaining calcium homeostasis and mitochondrial dynamic.
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Data availability
The data that support the findings of this study are available from the corresponding author upon reasonable request.
Abbreviations
- 3-MCPD:
-
3-Chloro-1, 2-propanediol
- SD:
-
Sprague-Dawley
- HEK293:
-
Human embryonic kidney
- LOVEL:
-
Lowest observed effect
- TDI:
-
Tolerable daily intake
- ER:
-
Endoplasmic reticulum
- UPR:
-
Unfolded protein response
- CKD:
-
Chronic kidney disease
- MAM:
-
Mitochondria-associated membranes
- 4-PBA:
-
4-Phenylbutyric acid
- H&E:
-
Hematoxylin and eosin
- DMSO:
-
Dimethyl sulfoxide
- MtMP:
-
Mitochondria membrane potential
- FBS:
-
Fetal bovine serum
- TEM:
-
Transmission electron microscope
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This work was supported by grants from National Natural Science Foundation of China (NSFC, 31571928), the Science and Technology Plan Project of Shaanxi Province (2020NY-106), and Student’s Platform for Innovation and Entrepreneurship Training Program (1201710712149, 1201710712127).
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Xiaoli Peng, Yujie Zhong, and Xiaodong Xia contributed to the conception and design of the experiment. Material preparation, data collection, and analysis were performed by Yujie Zhong, Chengni Jin, and Jiahui Han. The first draft of the manuscript was written by Yujie Zhong. Qi Liu, Dianjun Sun, and Jiachang Zhu commented on previous versions of the manuscript. All authors have read and approved the final manuscript.
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All animal procedures were performed in accordance with the Guide for the Care and Use of Laboratory Animals: Eighth Edition (ISBN-10: 0-309-15396-4) and approved by the Animal Ethics Committee of Northwest A&F University, and Chengdu Dossy Experimental Animals Co., Ltd (N19100398).
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Zhong, Y., Jin, C., Han, J. et al. Inhibition of ER stress attenuates kidney injury and apoptosis induced by 3-MCPD via regulating mitochondrial fission/fusion and Ca2+ homeostasis. Cell Biol Toxicol 37, 795–809 (2021). https://doi.org/10.1007/s10565-021-09589-x
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DOI: https://doi.org/10.1007/s10565-021-09589-x